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(a)explain how to measure k and resolution.

(b)state three method for measuring tmin reversed-phase chromatography.

(c)state three method for measuring tmin hydrophilic interaction liquid chromatography.

(d)Estimate tmfor 15×0.46column containing 5-μ³¾particles operating at a flow rate of 1.5mL/minEstimate tmif the particle size were 3.5-μ³¾ instead.

Short Answer

Expert verified

To measure k and resolution. state three method for measuring in reversed-phase chromatography. state three method for measuring in hydrophilic interaction liquid chromatography. Estimate particles operating at a flow.

a) Then we calculate using the formula:

k=tr-tmtm

b) The three methods for measuringtm in reversed-phase chromatography are:

measuring tmby running the unretained solutes uracil orNaNO3

tmis the time when the first baseline disturbance is observed we can use formula

tm=L.d2c2F.

c) tmcan be measured using toluene (detected at 215nm)

tmis the time when the first baseline disturbance is observed we can use formula:

tm=L.d2c2F

d)tm=1.1min

Step by step solution

01

definition of measure k

We can measure by measuring the retention time.

a) how to measure k and resolution.

We can measure by measuring the retention time for the peak of interest (tr)and the elution time for an unretained solute (tm) .

Then we calculate using the formula:

k=tr-tmtm

The resolution between neighboring peaks can be calculated dividing differences in their retention time by their average width at the baseline

resolution=Δ³Ùrwav

b)state three method for measuring

The three methods for measuringtm in reversed-phase chromatography are:

measuring tmby running the unretained solutes uracil orNaNO3

tmis the time when the first baseline disturbance is observed we can use formula

tm=L.d2c2F

Where the L is column lengthdc , is column diameter, and F is flow rate.

02

state three method for measuring

c)state three method for measuring

The three methods for measuringtmin hydrophilic interaction liquid chromatography are:

tmcan be measured using toluene (detected at 215nm)

tmis the time when the first baseline disturbance is observed we can use formula:

tm=L.d2c2F

d ) estimate column counting

The for tmμ³¾particle is:

tm=15cm.(0.46cm)22.1.5mL/mintm=1.1min

The for 3.5μ³¾particle is the same because tmdoes not depends on particle size.

Hence,

a) We can measure by measuring the retention time for the peak of interest (tr)and the elution time for an unretained solutelocalid="1655035932587" (tm).

Then we calculate using the formula:

k=tr-tmtm

b) The three methods for measuring tmin reversed-phase chromatography are:

measuring tmby running the unretained solutes uracil orNaNO3

tmis the time when the first baseline disturbance is observed we can use formula

tm=L.d2c2F

Where the L is column lengthdc, is column diameter, and F is flow rate.

c) The three methods for measuringtmin hydrophilic interaction liquid chromatography are:

tmcan be measured using toluene (detected at 215nm)

tmis the time when the first baseline disturbance is observed we can use formula:

tm=L.d2c2F

d)tm=1.1min

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Most popular questions from this chapter

A known mixture of compounds A and B gave the following HPLC results:

A solution was prepared by mixing 12.49mgof Bplus 10.00mLof unknown containing just and diluting to 25.00mL. Peak areas of 5.97and 6.38were observed for AandB, respectively. Find the concentration of A(mg/mL)in the unknown.

(a). Sketch a graph of the van Deemnter equation (plate height versus linear flow rate).what would the curve look like if the multiple path term were 0? If the longtitundinal diffusinal diffusion term were 0?

(b). Explain why the van Deemter curve for 1.8μ³¾particles in figure 25-3is nearly flat at high flow rate.what can you say about each of the terms in the van Deemeter equation for 1.8μ³¾particles.

(c). Explain why the 2.7μ³¾superficially porous particle enables separations similar to those achieved by 1.8μ³¾totally porous particles,but the superficially porous particle requires lower pressure.

  1. Use equation 25-1to estimate the length of a column required to achieve1.0×104plates if the stationary phase particles size is10.5,5.0,3.0,or1.5μ³¾

  2. If the retention time was 20mins on the 10.0μ³¾ particle size column, what is the retention time on the 5.0,3.0,or1.5μ³¾columns from part (a)? Assume that flow rate is constant for all columns.

  3. Use equation25-2to estimate the pressure of the column in (a) given that the pressure of the10.0μ³¾column was4.4Mpa

  4. If the flow rate was2.0mL/min , what is the baseline width for the peaks on 10.5,5.0,3.0,or1.5μ³¾columns form part (a)?

  5. Which of these column configurations would require a UHPLC instrument?

HPLC peak should generally not have an asymmetry factor, B/A in figure 23-14,outside the range0.9-1.5

  1. Sketch the shape of a peak with an asymmetry of 1.8
  2. What might you do to correct the asymmetry?

What are the general steps in developing as isocratic separation for reversed-phase chromatography with one organic solvent and temperature as variable?

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